Change search
ReferencesLink to record
Permanent link

Direct link
Subtracted Approaches to Gene Expression Analysis in Atherosclerosis
KTH, Superseded Departments, Biotechnology.
2003 (English)Doctoral thesis, comprehensive summary (Other scientific)
Abstract [en]

Gene expression analysis has evolved as an extensive toolfor elucidation of various biological and molecular eventsoccurring in different organisms. A variety of techniques andsoftware tools have been developed to enable easier and morerapid means of exploring the genetic information. A moreeffective approach than exploring the whole content of genesexpressed under certain conditions is to study fingerprintassays or to use subtracted cDNA libraries to identify onlydifferentially expressed genes.

The objective for the work in this thesis has been toexplore differentially expressed genes in atherosclerosis. Thiswas done by applying and modifying a protocol for thesubtractive approach RDA (Representational Difference Analysis)in different model systems.

Initially, the molecular effects of an anti-atheroscleroticdrug candidate were elucidated. In addition, two alternativeapproaches to identify differentially expressed genes obtainedafter iterative rounds of RDA subtraction cycles wereevaluated. This revealed that in most cases, the shotgunapproach in which the obtained gene fragments are clonedwithout any prior selection has clear advantages compared tothe more commonly used selection strategy, whereby distinctbands are excised after gel electrophoresis.

A key process in the atherosclerotic plaque initiation isthe phenotypic change of macrophages into foam cells, which canbe triggered in a model system by using macrophages exposed tooxidised LDL. To investigate the genes expressed in thisprocess, the RDA technique was combined with microarrayanalysis, which allows for selectivity and sensitivity throughRDA, as well as rapid high-throughput analysis usingmicroarrays. The combination of these techniques enablessignificant differences in gene expression to be detected, evenfor weakly expressed genes and the results to be reliablyvalidated in a high throughput manner.

Finally, investigation of the focal nature ofatherosclerotic lesions and gene expression profiling werestudied using in vivo aortic tissues from ApoE-/- and LDLR -/-mice. The study was based on a comparison between localisationsthat are likely, and others that are unlikely, to developatherosclerotic plaques, and the RDA technique was employed toexplore differential gene expression.

Keywords:Representational Difference Analysis,atherosclerosis, gene expression profiling

Place, publisher, year, edition, pages
Stockholm: Bioteknologi , 2003. , 61 p.
Keyword [en]
Representational Difference Analysis, atherosclerosis, gene expression profiling
URN: urn:nbn:se:kth:diva-3684ISBN: 91-7283-653-9OAI: diva2:9521
Public defence
NR 20140805Available from: 2004-01-22 Created: 2004-01-22Bibliographically approved

Open Access in DiVA

fulltext(2627 kB)828 downloads
File information
File name FULLTEXT01.pdfFile size 2627 kBChecksum MD5
Type fulltextMimetype application/pdf

By organisation

Search outside of DiVA

GoogleGoogle Scholar
Total: 828 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 243 hits
ReferencesLink to record
Permanent link

Direct link